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Circulation Research. 2001;89:891-898
Published online before print October 11, 2001, doi: 10.1161/hh2201.100205
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(Circulation Research. 2001;89:891.)
© 2001 American Heart Association, Inc.

Cellular Biology

Mitochondrial ATP-Sensitive Potassium Channels Attenuate Matrix Ca2+ Overload During Simulated Ischemia and Reperfusion

Possible Mechanism of Cardioprotection

Mitsushige Murata, Masaharu Akao, Brian O’Rourke, Eduardo Marbán

From the Institute of Molecular Cardiobiology, The Johns Hopkins University, Baltimore, Md.

Correspondence to Eduardo Marbán, MD, PhD, Institute of Molecular Cardiobiology, The Johns Hopkins University, 720 Rutland Ave Ross 844, Baltimore, MD 21205. E-mail marban{at}jhmi.edu

Abstract—— Mitochondrial ATP-sensitive potassium (mitoKATP) channels play a key role in ischemic preconditioning of the heart. However, the mechanism of cardioprotection remains controversial. We measured rhod-2 fluorescence in adult rabbit ventricular cardiomyocytes as an index of mitochondrial matrix Ca2+ concentration ([Ca2+]m), using time-lapse confocal microscopy. To simulate ischemia and reperfusion (I/R), cells were exposed to metabolic inhibition (50 minutes) followed by washout with control solution. Rhod-2 fluorescence gradually increased during simulated ischemia and rose even further with reperfusion. The mitoKATP channel opener diazoxide attenuated the accumulation of [Ca2+]m during simulated I/R (EC50=18 µmol/L). These effects of diazoxide were blocked by the mitoKATP channel antagonist 5-hydroxydecanoate (5HD). In contrast, inhibitors of the mitochondrial permeability transition (MPT), cyclosporin A and bongkrekic acid, did not alter [Ca2+]m accumulation during ischemia, but markedly suppressed the surge in rhod-2 fluorescence during reperfusion. Measurements of mitochondrial membrane potential, {Delta}{Psi}m, in permeabilized myocytes revealed that diazoxide depolarized {Delta}{Psi}m (by 12% at 10 µmol/L, P<0.01) in a 5HD-inhibitable manner. Our data support the hypothesis that attenuation of mitochondrial Ca2+ overload, as a consequence of partial mitochondrial membrane depolarization by mitoKATP channels, underlies cardioprotection. Furthermore, mitoKATP channels and the MPT differentially affect mitochondrial calcium homeostasis: mitoKATP channels suppress calcium accumulation during I/R, while the MPT comes into play only upon reperfusion.


Key Words: mitochondrial calcium overload • cardioprotection • ischemia




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